WO1998048027A2 - Produits agissant contre des affections retiniennes et traitement correspondant - Google Patents

Produits agissant contre des affections retiniennes et traitement correspondant

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Publication number
WO1998048027A2
WO1998048027A2 PCT/US1998/008003 US9808003W WO9848027A2 WO 1998048027 A2 WO1998048027 A2 WO 1998048027A2 US 9808003 W US9808003 W US 9808003W WO 9848027 A2 WO9848027 A2 WO 9848027A2
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Prior art keywords
promoter
cells
construct
raav
retinal
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PCT/US1998/008003
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English (en)
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WO1998048027A9 (fr
WO1998048027A3 (fr
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William W. Hauswirth
John G. Flannery
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University Of Florida
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Priority to EP98922042A priority Critical patent/EP0977880A2/fr
Priority to AU74675/98A priority patent/AU7467598A/en
Priority to BR9809284-7A priority patent/BR9809284A/pt
Priority to CA002287495A priority patent/CA2287495A1/fr
Priority to JP54626898A priority patent/JP2001527399A/ja
Publication of WO1998048027A2 publication Critical patent/WO1998048027A2/fr
Publication of WO1998048027A3 publication Critical patent/WO1998048027A3/fr
Publication of WO1998048027A9 publication Critical patent/WO1998048027A9/fr

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    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1136—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against growth factors, growth regulators, cytokines, lymphokines or hormones
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    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1137—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against enzymes
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    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1138—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against receptors or cell surface proteins
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • C12N2310/00—Structure or type of the nucleic acid
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    • C12N2310/121—Hammerhead
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    • C12N2310/00—Structure or type of the nucleic acid
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    • C12N2799/00—Uses of viruses
    • C12N2799/02—Uses of viruses as vector
    • C12N2799/021—Uses of viruses as vector for the expression of a heterologous nucleic acid
    • C12N2799/025—Uses of viruses as vector for the expression of a heterologous nucleic acid where the vector is derived from a parvovirus

Definitions

  • the subject invention was made with government support under a research project supported by NIH Grant Nos. EY07864 and EY11123 .
  • the government has certain ⁇ ghts m this invention.
  • Macular degeneration is a detenoration of the macula (the cone-rich center of vision) leading to gradual loss of central vision. Eventual loss of these cones leads to central vision loss and functional blindness. Macular degeneration may also have a genetic etiology or predisposition. Although a genetic etiology has not yet been established, based on pedigree studies it is very likely to exist. At least 500,000 individuals are estimated to suffer from macular degeneration currently in the United States.
  • the subject invention concerns materials and methods for achieving expression of proteins in retina cells.
  • the expression of proteins in retina cells can be used, for example, for the treatment of retinal diseases. More specifically, the subject invention provides polynucleotide sequences, and methods for using these sequences, to achieve highly specific expression of proteins in the retma. As described herein, the expression of these proteins can be used to treat a variety of retinal diseases.
  • the mate ⁇ als and methods of the subject invention can be used to treat autosomal dominant retmitis pigmentosa (ADRP).
  • ADRP autosomal dominant retmitis pigmentosa
  • the subject invention provides techniques for obtaining targeted, high level expression of any desired gene in the photoreceptor cells of the retina.
  • ribozymes can be highly and specifically expressed in the retina.
  • the ribozymes cleave the mutant forms of messenger RNA (mRNA) occurring in common forms of inherited retinal degeneration. This specificity makes these ⁇ bozymes able to destroy harmful mRNA while leaving normal mRNA mtact.
  • Ribozymes against other genetic forms of retmitis pigmentosa can be produced and used according to the subject invention.
  • Other polynucleotides encoding therapeutically useful products can also be selectively expressed in the eye using the teachings of the subject invention.
  • Figures la and lb show the construction of plasmids used according to the subject invention. These figures show a schematic diagram of the plasmid DNA constructs used to make rAAV viruses mOp-/ cZ (a) and Op-gfp (b).
  • TR 145 bp AAV terminal repeat sequence, mOp, 472 bp mu ⁇ ne rod opsm regulatory sequence from +86 to -388, SD/SA, 180 bp SV40 late viral protein gene 16S/19S splice donor and acceptor signal, lacZ; coding sequence for the bacterial lacZ gene; gfp, coding sequence for the synthetic green fluorescence gene; pA, pAl and pA2, polyadenylation signals; Epo, a tandem repeat of the polyoma virus enhancer region (bases 5210-5274); Ptk, thymidme kinase promoter of herpesvirus (bases 92-218); neo r , coding sequence of the neomycm resistance gene, Tn5 (bases 1555-2347) (Zolotukhin, S., M. Potter, W. Hauswirth, J. Guy, N. Muzyczka (1996) J. Virol. 70:4646-4654).
  • Ribozymes were: Hpl 1 hairpm ⁇ bozyme (filled circles), Hhl3 hammerhead ⁇ bozyme (filled triangles), Hpl "inactive” hairpin ⁇ bozyme (open circles), Hhl3 ⁇ "inactive” hammerhead ⁇ bozyme (open t ⁇ angle), or BOPS-g/p (X), all regulated by the same bovine opsm promoter. All injections were performed at P 14-15. The error bars were omitted if they fell withm the symbol, except for Hpl li at P75 and P90, where only one eye at each point was examined.
  • Rats Measurements of ONL thickness along the vertical me ⁇ dian of the eye from the optic nerve head (ONH) to the ora serrata (anterior margin of the retma) in rats at P90. Rats were either unmjected (open triangles) or injected at P14-15 with Hpl 1 hairpm ribozymes (filled circles) or Hhl3 hammerhead ribozymes (open circles).
  • the subject invention pertains to materials and methods for achieving highly specific expression of desired proteins in the retina. These proteins can be used as described herein to achieve a beneficial therapeutic affect.
  • the subject invention provides materials and methods which can be used to reduce or eliminate the symptoms of mhe ⁇ ted eye disease caused by mutations in genes for retinal proteins.
  • the subject invention provides materials and methods for achieving efficient and cell type-specific expression of exogenous genes in photoreceptor cells (PRs) of the mammalian retina.
  • PRs photoreceptor cells
  • rAAV recombmant Adeno-associated Virus
  • the subject invention provides a method for treating autosomal dominant retmitis pigmentosa (ADRP) at a molecular level.
  • ADRP autosomal dominant retmitis pigmentosa
  • Gene therapy for ADRP involves (1) an efficient and cell-type specific gene delivery/expression system, and (2) a selective means of inhibiting production of the mutant protein.
  • rAAV vectors are used to transfer the bacte ⁇ al lac gene or a synthetic green fluorescent protein gene (gfp) to mouse or rat retinas following injection into the subretmal space.
  • gfp synthetic green fluorescent protein gene
  • results demonstrate the surprising and advantageous ability to achieve highly specific expression of proteins m the mammalian retma.
  • reporter gene product was found exclusively in photoreceptors, not in any other retinal cell type or in the adjacent retinal pigment epithelium (RPE).
  • GFP-expressmg photoreceptors typically encompassed 10-20% of the total retinal area following a single 2 ⁇ injection. Both rod and cone photoreceptors were transduced with nearly 100% efficiency in the region directly surrounding the injection site. Approximately 2.5 million photoreceptors were transduced as a result of the single subretmal inoculation. The use of such proximal opsm promoters therefore can be used to substantially enhance the efficiency of expression in cases where rod and/or cone -specific expression of a potentially therapeutic gene is desired.
  • the gfp- contaimng rAAV stock was substantially free of both adenovirus and wild-type AAV, as judged by plaque assay and infectious center assay, respectively.
  • helper virus- free rAAV vectors can achieve high frequency tissue-specific transduction of terminally differentiated, postmitotic photoreceptor cells.
  • the human rod opsm, or cone opsm, promoter analogous to the mouse sequence would be used.
  • the appropriate regions of the human sequence can be readily identified and used by the skilled artisan having the benefit of the instant disclosure Assuming a 20 ⁇ l injection into the central human retma at the region of greatest rod density, approximately 8 million rods would be expected to be transduced m a focal region encompassing approximately 6% of the total retina. By transducing such an area of PRs it is possible to improve or delay retinal degenerations in a variety of mhe ⁇ ted retinal diseases.
  • the mOp-gfp- rAAV plasmid DNA was made by first adding Not I linkers to the 472bp Bgl II/Xho I proximal opsm promoter fragment of pRG3 and inserting it into the Not I sites of pTRUF2 (Zolotukhin, S. M. Potter, W. Hauswirth, J. Guy, N. Muzyczka (1996) J Virol. 70:4646-4654) (Fig. lb) rAAV virus production and analysis.
  • human 293 cells were co-transfected with mOp-/ ⁇ cZ-rAAV or mOp-g/ >-rAAV plasmid DNA and the helper pIM45 plasmid DNA carrying the wtAAV genome without terminal repeats (Zolotukhm, S. M Potter, W. Hauswirth, J. Guy, N. Muzyczka (1996) J. Virol.
  • the RT-PCR employed a two buffer thermostable Tth polymerase system (Promega, Madison, WI) according to manufacturer's instructions and lacL sequence p ⁇ mers from nucleotides 105 to 124 (forward) and 303 to 286 (reverse). Rnase and Dnase digestions prior to the RT-PCR were performed as previously desc ⁇ bed (van Gmkel, P., W. Hauswirth (1994) J. Biol. Chem. 269:4986-4992).
  • Example 1 Design of rAAV Vectors for Gene Transfer to Photoreceptors
  • a 472bp of the proximal murme rod opsm promoter (+86 to -385) was linked to a lacZ-SV40 polyA reporter gene and then inserted this into pTR.
  • the gene construct was packaged into
  • AAV virus particles concentrated, tested for contaminating Adenovirus and titered for recombmant AAV by an infectious center assay.
  • the ⁇ ght eyes of 30 C57B1/6J mice were injected sub-retmally with l ⁇ l of mOp-/ ⁇ cZ virus (10 7 m per ml). After two weeks, the right
  • test and left (control) eyes of 12 animals were removed, fixed and stained with X-gal.
  • Test retina in 6 of 12 injected eyes exhibited a focal blue region consistent with a subretmal bleb of the injected virus creating a localized retinal detachment. All control eyes showed no X-gal reaction. Reporter gene expression was examined in mice sacrificed at later periods and was detected at 10 weeks post-mjection suggesting persistent reporter transgene expression.
  • lacZ gene product was analyzed at higher resolution by preparing se ⁇ al 50 ⁇ m transverse sections from the entire whole mounts.
  • the blue X-gal reaction product is observed p ⁇ manly in the PR inner segments.
  • Most of the PRs were filled with X-gal in this region.
  • X-gal staining was slightly above control levels in the PR synaptic termini m the outer plexiform layer.
  • PR outer segments, RPE and other retinal cells in this region did not reveal X- gal staining above baseline levels observed m identically treated, unmjected or PBS-mjected control returns from the contralateral eye.
  • GFP fluorescence typically extended over 10-20% of the retinal area m a radial pattern from the injection site. Immediately surrounding the point of infection, the transduction frequency, as judged by the intensity of GFP fluorescence, was very high, with a continuous positive signal. In transverse sections extending from the central retma to the pe ⁇ phery, beyond a region of apparently saturated GFP fluorescence, the percentage of transduced cells decreased radially with distance from the injection site. GFP-positive cells were easily identifiable as PRs by their specialized shape and location m the retma. Hence, only PR cells appeared to have been transduced, i.e., infected by the rAAV and expressing the gfp passenger gene.
  • Example 3 Opsm Promoter Confers Photoreceptor Cell Specificity
  • the PR-specific pattern of GFP expression was confirmed by laser confocal microscopy. GFP was not observed between the inner limiting membrane (vitreal face of the inner retma) and the outer plexiform layer (OPL) (junction of the inner retma with PR synaptic termini). This region contains all the non-PR retinal neuronal (bipolar, ho ⁇ zontal, amac ⁇ ne, and ganglion) and ghal (M ⁇ ller) cells. Virtually 100% of the PR inner segments, cell bodies, and synaptic terminals exhibited strong GFP fluorescence. In regions more peripheral to the injection site, the fraction of positive PRs was substantially reduced, consistent with the radial decline in fluorescence seen retinal whole mounts. It was established that all PR cell bodies contained
  • GFP signal by examining se ⁇ al optical sections (0.32 ⁇ m). Through-focus series demonstrated that occasional, dark regions in the ONL always contained a gfp-positive PR cell body in another plane of section. Therefore, all PRs, including both rods and cones, supported reporter gene expression. Outer segments demonstrated less fluorescence than other PR compartments, near the level of autofluorescence seen in control outer segments. No GFP signal was observed m the REP, choroid, or sclera.
  • GFP-positive PRs The area of GFP-positive PRs resulting from a typical injection from epifluorescence images of retinal whole mounts was established. GFP -positive areas were measured with NIH
  • Image software by segmenting the image into regions of GFP fluorescence and background on the basis of gray level. Area measurements were calibrated by imaging a 1000 ⁇ m reference scale on the film together with the whole mount.
  • the retinal area that contained 50% or more PR cells positive for GFP signal m whole mounts was measured. On average, the GFP-positive area covered -35% of the total retinal area of the rat retma.
  • the number of GFP -positive PRs resulting from a typical injection was estimated by examining se ⁇ al optical sections taken through the retma. Serial confocal images suggest 100% PR transduction m the region directly adjacent to the injection site, since we did not observe GFP-negative cell bodies within the outer nuclear layer m adjacent confocal optical sections. It is estimated that the whole rat retina contains 15.7 million PR cells. From these observations, a conservative estimate is that 2-3 million PRs were transduced by the gfp- containmg rAAV.
  • Example 5 Construction of Vectors and Expression in Target Cells rAAV- ⁇ bozyme constructs. Recombmant AAV constructs were based on the pTR-UF2 vector (Zolotukhm, S., M. Potter, W.W. Hauswirth et al. [1996] J Virol. 70:4646-4654). They resemble the vector used by Flannery et al. (Flannery, J.G., S. Zolotukhm, M.I. Vaquero et al [1997] Proc. Natl.
  • bovine promoter fragment contains three proximal promoter elements and the endogenous transc ⁇ ptional start site at its 3' end (DesJardm, L.E., W W. Hauswirth [1996] Inv Ophth. Vis. Sci. 37:154-165) and supports high efficiency, rat photoreceptor-specific expression in vivo.
  • ⁇ bozymes Active and inactive ⁇ bozymes were designed, tested and cloned. Each ⁇ bozyme gene was followed by an internally cleaving hairpm nbozyme derived from plasmid pHC (Althoffr, M., R. T ⁇ tz, A.A. Hampel [1992] Gene 122:85-90) resulting in ⁇ bozyme cassettes of 140-152 bp. Self cleavage at the internal cutting site m the primary nbozyme RNA leaves identical 3' ends on each mature nbozyme.
  • the ⁇ bozyme cassette was preceded by an intron derived from SV40 and followed by a polyadenylation signal in order to promote nuclear export of the nbozyme.
  • Recombmant AAV titers were determined using both an infectious center assay (Flannery, J.G., Zolotukhm, S. Vaquero et al. [1997] Proc. Natl. Acad. Sci. USA 94:6916-6921) and a DNAse resistant physical particle assay employing a quantitative, competitive PCR of the ne gene contained within all rAAV- ⁇ bozyme particles (Zolotukhm, S., M. Potter, W.W. Hauswirth et al. [1996]
  • Each of the four rAAV- ⁇ bozyme virus preparations contained 10 10 to 10" DNASE resistant particles per ml and 10 8 to 10 9 infectious center units per ml. Contaminating helper adenovirus and wild-type AAV, assayed by serial dilution cytopathic effect or infectious center assay respectively, were less than five order of magnitude lower than rAAV.
  • RNA retma were isolated without fixation and total RNA immediately extracted using the
  • RNeasy Minikit Qiagen, Santa Clanta, CA. RT-PCR was performed using the Pharmacia First-Strand cDNA synthesis kit employing ohgo dT as the primer. Wild-type and transgene opsm cDNAs were amplified using a three p ⁇ mer system desc ⁇ bed above. P ⁇ mers specific for ⁇ -actm cDNA (Timmers, A.M., B.R. Newton, W.W. Hauswirth [1993] Exp Eye Res 56:251- 265) were included in each reaction for internal standardization.
  • Ribozymes were designed to recognize and cleave the unique transc ⁇ pt produced by the P23H transgene.
  • the mutant target sequence "5 '-UCGGAGUCUACUUCG-3 '" (SEQ ID NO. 17) contains two differences from the wild-type mRNA (indicated in bold). The hairpm nbozyme (Hpl l) cleaved 3' to the first adenosine residue (underlined) and the hammerhead ⁇ bozyme (Hhl3) cleaved 3' to the central cytosme residue (underlined).
  • Control nbozymes (Hpl li and Hhl3 ⁇ , respectively) retained the targeting domains but contained fatal flaws in their catalytic domains.
  • the active hammerhead nbozyme (Hhl3) was able to cleave 20% of the P23H target within 10 mm. of incubation and by 5 hours greater than 80% was converted to the expected products.
  • both nbozymes exhibited kinetic constants (K,-, and k- at ) similar to those of naturally occurnng ⁇ bozymes.
  • the two active ribozymes produced negligible cleavage of the wild-type transcnpt even in the presence of high MgCl 2 concentrations.
  • Control ⁇ bozymes (Hpl li and Hhl3 ⁇ ) containing inactivating mutations in their catalytic domains were without measurable activity on any substrate Using total RNA denved from retinas of P23H rats on P62, both the hairpm and the hammerhead ⁇ bozymes were able to cleave the mRNA product of the mutant transgene selectively.
  • TgN(P23H)3 (abbreviated P23H-3), that has a retinal degeneration phenotype similar to patients with retmitis pigmentosa (Steinberg, R.H., J.G. Flannery, M.I. Naash et al. [1996] Inv. Ophth Vis. Sci 37:S698) was used.
  • mutated opsm transgene begins at about postnatal day (P) 5 in rats, leading to a gradual death of photoreceptor cells. These rats develop an apparently normal retma up to PI 5, although there are somewhat more pyknotic photoreceptor nuclei m the outer nuclear layer (ONL) than m non-transgemc control rats. Thereafter, death of photoreceptor cells is almost linear until about P60, resulting m loss of about 40% of the photoreceptors. After P60, the rate of cell loss decreases, until by one year the retinas have less than a single row of photoreceptor nuclei.
  • the rAAV-nbozyme vector was injected into the mterphotoreceptor space between the photoreceptors and the adjacent retinal pigment epithelium at P14 or P15. Rats were sacnficed and eyes examined at 3 time points between P60-P90. At these ages in unmjected control eyes of P23H-3 rats, the ONL thickness, which is an index of photoreceptor cells number, was reduced to about 60% of normal.
  • Ribozyme-mjected eyes showed a modest but significant decrease in the accumulation of transcnpt denved from the P23H transgene. Control eyes exhibited little vanation in the level of transgene opsm mRNA. Eyes injected with either active ⁇ bozyme uniformly exhibited lowered transgene mRNA levels relative to total opsm mRNA in the same eye. Retinas receiving the hairpm ⁇ bozyme Hpl 1 showed a 15.3 ⁇ 3.3% decrease m transgene expression, and those with the hammerhead nbozyme Hhl3 showed a decrease of 11.1+5.1% decrease
  • the upper bound denves from the DNAse resistant particle assay, indicating that 2 ⁇ l of the rAAV- ⁇ bozyme virus preparation contained 2 x 10 7 to 10 8 rAAV This is an upper bound because not all particles counted are expected to be infectious.
  • the lower bound for rAAV titer is generated by the infectious center assay, indicating 10 6 to 10 7 rAAV per ⁇ l.
  • rAAV retinal pigment epithelium
  • the initial volume of extracellular space approximately 0.5 ⁇ l, increases greatly with the 2 ⁇ l injection.
  • the fluid transport function of the RPE dehydrates this space, reapposmg the photoreceptors and RPE and concentrating the rAAV
  • the detachment of the photoreceptors from the RPE resolves withm several hours Du ⁇ ng the reattachment process, viral particles are spread laterally through the subretmal space.
  • RIS the unmjected control retinas had RIS that were about 90% of normal. Both the active and inactive ⁇ bozymes resulted m RIS lengths of 98% or greater of the normal length, and about 10- 15% longer than unmjected controls.

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Abstract

La présente invention concerne des produits et des traitements suscitant une expression satisfaisante et spécifique de protéines dans les cellules photorécepteurs de la rétine. Plus particulièrement, les produits de recombinaison sont constitués d'un vecteur viral associé aux adénovirus renfermant un promoteur d'opsine des bâtonnets ou des cônes rétiniens. Ces produits et traitement conviennent à des thérapies adaptées à des affections rétiniennes. Selon une réalisation, pour soigner la rétinite pigmentaire, on utilise des ribozymes qui dégradent l'ARNm mutant.
PCT/US1998/008003 1997-04-21 1998-04-21 Produits agissant contre des affections retiniennes et traitement correspondant WO1998048027A2 (fr)

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EP98922042A EP0977880A2 (fr) 1997-04-21 1998-04-21 Produits agissant contre des affections retiniennes et traitement correspondant
AU74675/98A AU7467598A (en) 1997-04-21 1998-04-21 Materials and methods for treatment of retinal diseases
BR9809284-7A BR9809284A (pt) 1997-04-21 1998-04-21 Construto e processo para expressão de polinucleotìdeo em altos nìveis
CA002287495A CA2287495A1 (fr) 1997-04-21 1998-04-21 Produits agissant contre des affections retiniennes et traitement correspondant
JP54626898A JP2001527399A (ja) 1997-04-21 1998-04-21 網膜疾患の処置のための材料および方法

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WO2001083729A2 (fr) * 2000-05-01 2001-11-08 Novartis Ag Vecteurs pour transduction oculaire et utilisation en therapie genique
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WO2007131180A2 (fr) 2006-05-04 2007-11-15 Wayne State University Restauration de réponses visuelles par administration in vivo d'acides nucléiques de rhodopsine
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US8147823B2 (en) 2001-04-13 2012-04-03 The Trustees Of The University Of Pennsylvania Method of treating or retarding the development of blindness
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US9180309B2 (en) 2010-02-26 2015-11-10 Cornell University Retina prosthesis
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US10947281B2 (en) 2012-03-05 2021-03-16 Wayne State University Identification of channelrhodopsin-2 (ChR2) mutations and methods of use
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US8551970B2 (en) 1996-04-02 2013-10-08 Optigen Patents Limited Genetic suppression and replacement
WO2004020631A2 (fr) * 1996-04-02 2004-03-11 Optigen Patents Limited Substitution et suppression genetique
WO2004020631A3 (fr) * 1996-04-02 2004-07-29 Optigen Technologies Ltd Substitution et suppression genetique
WO1998048009A3 (fr) * 1997-04-21 1999-03-25 Univ Florida Substances et methodes de traitement de maladies retiniennes par les ribozymes
US6225291B1 (en) 1997-04-21 2001-05-01 University Of Florida Rod opsin mRNA-specific ribozyme compositions and methods for the treatment of retinal diseases
WO1998048009A2 (fr) * 1997-04-21 1998-10-29 University Of Florida Substances et methodes de traitement de maladies retiniennes par les ribozymes
US7342111B2 (en) 1999-04-30 2008-03-11 University Of Florida Research Foundation, Inc. Adeno-associated virus-delivered ribozyme compositions and methods of use
WO2001083729A2 (fr) * 2000-05-01 2001-11-08 Novartis Ag Vecteurs pour transduction oculaire et utilisation en therapie genique
WO2001083729A3 (fr) * 2000-05-01 2002-09-19 Novartis Ag Vecteurs pour transduction oculaire et utilisation en therapie genique
US9433688B2 (en) 2001-04-13 2016-09-06 The Trustees Of The University Of Pennsylvania Method of treating or retarding the development of blindness
US8147823B2 (en) 2001-04-13 2012-04-03 The Trustees Of The University Of Pennsylvania Method of treating or retarding the development of blindness
US9730981B2 (en) 2006-05-04 2017-08-15 Salus University Restoration of visual responses by in vivo delivery of rhodopsin nucleic acids
EP3437473A1 (fr) 2006-05-04 2019-02-06 Wayne State University Restauration de réponses visuelles par administration in vivo d'acides nucléiques de la rhodopsine
WO2007131180A2 (fr) 2006-05-04 2007-11-15 Wayne State University Restauration de réponses visuelles par administration in vivo d'acides nucléiques de rhodopsine
US10426844B2 (en) 2008-05-20 2019-10-01 University Of Florida Research Foundation, Incorporated Capsid-mutated rAAV vectors and methods of use
US10039921B2 (en) 2010-02-26 2018-08-07 Cornell University Retina prosthesis
US9180309B2 (en) 2010-02-26 2015-11-10 Cornell University Retina prosthesis
US9220634B2 (en) 2010-02-26 2015-12-29 Cornell University Retina prosthesis
EP3088044A1 (fr) 2010-02-26 2016-11-02 Cornell University Prothèse de rétine
US10561841B2 (en) 2010-02-26 2020-02-18 Cornell University Retina prosthesis
EP3669933A1 (fr) 2010-02-26 2020-06-24 Cornell University Prothèse de rétine
US11771763B2 (en) 2010-04-05 2023-10-03 Eos Neuroscience, Inc. Methods and compositions for decreasing chronic pain
US9453241B2 (en) 2010-05-04 2016-09-27 Wayne State University AAV-mediated subcellular targeting of heterologous rhodopsins in retinal ganglion cells
US9968689B2 (en) 2010-05-04 2018-05-15 Wayne State University AAV-mediated subcellular targeting of heterologous rhodopsins in retinal ganglion cells
US9925373B2 (en) 2010-09-10 2018-03-27 Cornell University Neurological prosthesis
US9302103B1 (en) 2010-09-10 2016-04-05 Cornell University Neurological prosthesis
US11739294B2 (en) 2011-05-18 2023-08-29 The Regents Of The University Of California Compositions and methods for treating retinal diseases
US10781422B1 (en) 2011-05-18 2020-09-22 The Regents Of The University Of California Compositions and methods for treating retinal diseases
US10752882B2 (en) 2011-05-18 2020-08-25 The Regents Of The University Of California Compositions and methods for treating retinal diseases
US10822585B2 (en) 2011-05-18 2020-11-03 The Regents Of The University Of California Compositions and methods for treating retinal diseases
WO2013029008A1 (fr) 2011-08-25 2013-02-28 Cornell University Codeur rétinien pour vision industrielle
US10769483B2 (en) 2011-08-25 2020-09-08 Cornell University Retinal encoder for machine vision
US10303970B2 (en) 2011-08-25 2019-05-28 Cornell University Retinal encoder for machine vision
US9547804B2 (en) 2011-08-25 2017-01-17 Cornell University Retinal encoder for machine vision
US11640681B2 (en) 2011-08-25 2023-05-02 Cornell University Retinal encoder for machine vision
US9375491B2 (en) 2011-10-28 2016-06-28 University Of Florida Research Foundation, Inc. Chimeric promoter for cone photoreceptor targeted gene therapy
WO2013063601A1 (fr) * 2011-10-28 2013-05-02 University Of Florida Research Foundation, Inc. Promoteur chimérique pour thérapie génique ciblant un photorécepteur à cônes
US10947281B2 (en) 2012-03-05 2021-03-16 Wayne State University Identification of channelrhodopsin-2 (ChR2) mutations and methods of use
US10515269B2 (en) 2015-04-20 2019-12-24 Cornell University Machine vision with dimensional data reduction
US11430263B2 (en) 2015-04-20 2022-08-30 Cornell University Machine vision with dimensional data reduction
WO2020154535A1 (fr) * 2019-01-23 2020-07-30 University Of Florida Research Foundation, Incorporated Transduction et propagation latérale hautement efficaces dans la rétine par un nouveau virus aav amélioré par conception rationnelle

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CA2287495A1 (fr) 1998-10-29
JP2001527399A (ja) 2001-12-25
BR9809284A (pt) 2004-08-03
WO1998048027A9 (fr) 1999-04-29
EP0977880A2 (fr) 2000-02-09
WO1998048027A3 (fr) 1999-03-25
AU7467598A (en) 1998-11-13

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